• Journal of Microbiology and Biotechnology
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• v.29 no.5
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• pp.749-757
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• 2019

Nitrilase is a valuable hydrolase that catalyzes nitriles into carboxylic acid and ammonia. Its applications, however, are severely restricted by the harsh conditions of industrial reaction processes. To solve this problem, a nitrilase from Acidovorax facilis 72W was inserted into an Escherichia coli-Bacillus subtilis shuttle vector for spore surface display. Western blot, enzyme activity measurements and flow cytometric analysis results all indicated a successful spore surface display of the CotB-nit fusion protein. In addition, the optimal catalytic pH value and temperature of the displayed nitrilase were determined to be 7.0 and $50^{\circ}C$, respectively. Moreover, results of reusability tests revealed that 64% of the initial activity of the displayed nitrilase was still retained at the $10^{th}$ cycle. Furthermore, hydrolysis efficiency of upscale production of cyanocarboxylic acid was significantly higher in the displayed nitrilase-treated group than in the free group expressed by E. coli (pET-28a-nit). Generally, the display of A. facilis 72W nitrilase on the spore surface of Bacillus subtilis may be a useful method for immobilization of enzyme and consequent biocatalytic stabilization.

• Journal of Microbiology and Biotechnology
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• v.26 no.7
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• pp.1267-1277
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• 2016

Currently, enzymatic synthesis of lactulose, a synthetic prebiotic disaccharide, is commonly performed with glycosyl hydrolases. In this work, a new type of lactulose-producing biocatalyst was developed by displaying β-galactosidase from Bacillus stearothermophilus IAM11001 (Bs-β-Gal) on the surface of Bacillus subtilis 168 spores. Localization of β-Gal on the spore surface as a fusion to CotX was verified by western blot analysis, immunofluorescence microscopy, and flow cytometry. The optimum pH and temperature for the resulting spore-displayed β-Gal was 6.0 and 75℃, respectively. Under optimal conditions, it showed maximum activity of 0.42 U/mg spores (dry weight). Moreover, the spore-displayed CotX-β-Gal was employed as a whole cell biocatalyst to produce lactulose, yielding 8.8 g/l from 200 g/l lactose and 100 g/l fructose. Reusability tests showed that the spore-displayed CotX-β-Gal retained around 30.3% of its initial activity after eight successive conversion cycles. These results suggest that the CotX-mediated spore-displayed β-Gal may provide a promising strategy for lactulose production.

• 제어로봇시스템학회:학술대회논문집
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• 1987.10b
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• pp.249-254
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• 1987

This paper presents a new method for generating smooth free-form surface by local correction. B-spline surface is used for its convenience of local correction, and the direction of surface correction is fixed to the average-surface-normal direction. The surface to be corrected is approximated into a uniform cubic B-spline surface. Then, the smoothness (curvature arrows, iso-parametric lines) of the approximated surface is displayed with B-spline control points. When a control point near the region that needs correction is selected, a new point 1 mm higher than the original control point in the direction of the average surface normal is displayed. And the surface is corrected by giving the amount of control point movement interactively. Since the direction of correction is given by the program and the amount of correction is selected by the user, the method is called semiautomatic. sufficiently smooth surface can be obtained by this method. Examples are given to illustrate the method.

• Macromolecular Research
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• v.12 no.6
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• pp.553-558
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• 2004

An ultrafiltration membrane (polyethersulfone, PM 10) was surface-modified by treating it with low-tem­perature plasmas of oxygen, acrylic acid (AA), acetylene, diaminocyolohexane (DACH), and hexamethyldisiloxane (HMDSO). The effects that these modifications have on the filtration efficiency of a membrane in waste water treat­ment were investigated. The oxygen, AA, and DACH plasma-treated membranes became more hydrophilic. The water contact angles ranged from < $10^{\circ}\;to\;55^{\circ}$ depending on the type of plasma and the treatment conditions. The oxygen plasma-treated membranes displayed a higher initial flux $(312-429\%),$ but lower rejection $(6-91\%),$ than did an untreated membrane. The AA plasma-treated membranes displayed lower or higher initial flux $(42-156\%),$ depending upon the treatment conditions, but higher rejection $(224-295\%)$ in all cases. The DACH plasma-treated membranes displayed lower initial flux. All of them, especially the AA plasma-treated membrane, displayed improved fouling resistance with either a slower or no flux decline. Acetylene and HMDSO plasma-treated membranes became more hydrophobic and displayed both lower initial flux and lower fouling resistance.

• The Journal of Korea Robotics Society
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• v.8 no.3
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• pp.179-185
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• 2013

In this paper we propose a haptic interaction system that physically represents the underlying geometry of objects displayed in a 2D picture, i.e., a digital image. To obtain the object's geometry displayed in the picture, we estimate the physical transformation between the object plane and the image plane based on homographic information. We then calculate the rotated surface normal vector of the object's face and place it on the corresponding part in the 2D image. The purpose of this setup is to create a force that can be rendered along with the image without distorting the visual information. We evaluated the proposed haptic rendering system using a set of pictures of objects with different orientations. The experimental results show that the participants reliably identified the geometric configuration by touching the object in the picture. We conclude this paper with a set of applications.

• Journal of the Society of Naval Architects of Korea
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• v.29 no.3
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• pp.15-20
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• 1992

This paper outlines the methods of visualizing 3-dimensional free form surfaces employing the Painter's algorithm, especially for the ship hull forms which are defined as open uniform Bi-cubic B-spline surfaces. The computer graphic codes are developed for the transparent wire-frame, the hidden surface removal and the shading visualization techniques, The codes are applied to the ship hull 3-dimensional surface visualization and the color graphic figures are displayed. Also Gaussian curvature is displayed on the color plots of the isoparametric net of the ship hull surface.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.468-472
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• 2003

Recently, genetic engineering techniques have been used to display various heterologous peptides and proteins (enzyme, antibody, antigen, receptor and fluorescence protein, etc.) on the yeast cell surface. Living cells displaying various enzymes on their surface could be used repeatedly as 'whole cell biocatalysts' like immobilized enzymes. We constructed a yeast based whole cell biocatalyst displaying T. reesei cellobiohydrolase I (CBH I ) on the cell surface and endowed the yeast-cells with the ability to degrade cellulose. By using a cell surface engineering system based on ${\alpha}-agglutinin,$ CBH I was displayed on the cell surface as a fusion protein containing the N-terminal leader peptide encoding a Gly-Ser linker and the $Xpress^{TM}$ epitope. Localization of the fusion protein on the cell surface was confirmed by confocal microscopy. In this study, we report on the genetic immobilization of T. reesei CBH I on the S. cerevisiae and hydrolytic activity of cell surface displayed CBH I.

• Journal of Microbiology and Biotechnology
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• v.24 no.9
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• pp.1178-1188
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• 2014

In this study, the yeast Pichia pastoris was genetically modified to assemble minicellulosomes on its cell surface by the heterologous expression of a truncated scaffoldin CipA from Clostridium acetobutylicum. Fluorescence microscopy and western blot analysis confirmed that CipA was targeted to the yeast cell surface and that NtEGD, the Nasutitermes takasagoensis endoglucanase that was fused with dockerin, interacted with CipA on the yeast cell surface, suggesting that the cohesin and dockerin domains and cellulose-binding module of C. acetobutylicum were functional in the yeasts. The enzymatic activities of the cellulases in the minicellulosomes that were displayed on the yeast cell surfaces increased dramatically following interaction with the cohesin-dockerin domains. Additionally, the hydrolysis efficiencies of NtEGD for carboxymethyl cellulose, microcrystal cellulose, and filter paper increased up to 1.4-fold, 2.0-fold, and 3.2-fold, respectively. To the best of our knowledge, this is the first report describing the expression of C. acetobutylicum minicellulosomes in yeast and the incorporation of animal cellulases into cellulosomes. This strategy of heterologous cellulase incorporation lends novel insight into the process of cellulosome assembly. Potentially, the surface display of cellulosomes, such as that reported in this study, may be utilized in the engineering of S. cerevisiae for ethanol production from cellulose and additional future applications.

• Journal of Microbiology and Biotechnology
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• v.15 no.1
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• pp.141-146
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• 2005

A new system for displaying proteins on the surface of Escherichia coli was developed using the Salmonella typhimurium outer membrane protein C (OmpC) as an anchoring motif. The C-terminal deletionfusion strategy was developed to fuse the polyhistidine peptides and green fluorescent protein (GFP) to the Cterminal of the truncated functional portion of OmpC. The polyhistidine peptides of up to 243 amino acids could besuccessfully displayed on the E. coli cell surface, which allowed recombinant E. coli to adsorb up to 34.2 μmol of Cd2+ per gram dry cell weight. The GFP could also be successfully displayed on the E. coli cell surface. These results suggest that the C-terminal deletion-fusion strategy employing the S. typhimurium OmpC as an anchoring motif provides a new efficient way for the display of large proteins on the surface of E. coli.

• Journal of Microbiology and Biotechnology
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• v.18 no.5
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• pp.946-951
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• 2008

The opd gene, encoding organophosphorus hydrolase (OPH) from Flavobacterium sp. capable of degrading a wide range of organophosphate pesticides, was surface- and intracellular-expressed in Synechococcus PCC7942, a prime example of photoautotrophic cyanobacteria. OPH was displayed on the cyanobacterial cell surface using the truncated ice nucleation protein as an anchoring motif. A minor fraction of OPH was displayed onto the outermost surface of cyanobacterial cells, as verified by immunostaining visualized under confocal laser scanning microscopy and OPH activity analysis; however, a substantial fraction of OPH was buried in the cell wall, as demonstrated by proteinase K and lysozyme treatments. The cyanobacterial outer membrane acts as a substrate (paraoxon) diffusion barrier affecting whole-cell biodegradation efficiency. After freeze-thaw treatment, permeabilized whole cells with intracellular-expressed OPH exhibited 14-fold higher bioconversion efficiency ($V_{max}/K_m$) than that of cells with surface-expressed OPH. As cyanobacteria have simple growth requirements and are inexpensive to maintain, expression of OPH in cyanobacteria may lead to the development of a low-cost and low-maintenance biocatalyst that is useful for detoxification of organophosphate pesticides.